Alterations of enzymatic activities during red and white muscle differentiation in vitro

The capacity of embryonic myoblasts, obtained from muscles which are known to be composed in the adult chicken of only one type of fibre (white type IIB or red type I) to give rise to fibres with similar or distinctive characteristics has been investigated in vitro. The specific activities of several enzymes related to the principal metabolic pathways were assayed and, in addition, the activities of CPK and AchE were determined on muscle cell cultures 2, 4, 6, 8, 10, 14 and 21 days old. The evidence presented indicates that both ‘red’ and ‘white’ myogenic cells appear to start out the same way, but differ later after prolonged incubation (21 days). This suggests differences in enzymatic properties between the two cellular types which will develop later.     

Effect of lumbar extensor fatigue on paraspinal muscle reflexes

Low back disorders are a frequent medical problem. Altered neuromuscular control of the spine has been associated with low back pain, and may contribute to its occurrence. The purpose of this study was to investigate the effect of lumbar extensor fatigue on reflex delay and amplitude in the paraspinal muscles. Ten healthy males (20–22 years of age) were subjected to an anteriorly-directed perturbation applied at the inferior margin of the scapulae while standing quietly before and after a lumbar extensor fatiguing protocol. The fatiguing protocol consisted of multiple sets of back extensions and intermittent isometric maximum voluntary contraction on a Roman chair for 14 min until 60% of unfatigued lumbar extensor MVC was reached. Reflexes were recorded from the paraspinal muscles at the level of L4. Results indicated the mean reflex delay was 60 ± 18 ms and was not affected by fatigue (p = 0.278). Reflex amplitude increased 36 ± 32% with fatigue (p = 0.017). The increase in reflex amplitude may reflect an attempt to compensate for losses in muscle force capacity with fatigue in order to maintain sufficient spinal stability. However, additional studies are necessary to investigate the mechanisms of this fatigue-related change in paraspinal reflex.     

Determination of the polymorphic composition of smooth pea starch

A method for estimating the proportions of ‘A’ and ‘B’ polymorphs comprising a sample of ‘C’ type starch is proposed which uses established experimental techniques with commercially available spreadsheet and X-ray analysis software. Waxy maize, potato and smooth pea starches were used to provide X-ray diffraction patterns characteristic of the ‘A’, ‘B’ and ‘C’ starch polymorphs. Samples of amorphous starches were also prepared. The method initially involved subtraction of the amorphous phase and instrumental background from the X-ray diffraction patterns of each starch sample using the spreadsheet program, Lotus 1-2-3. The remainder of the pattern, representing the crystalline portion of the starch sample, was then analysed by profile fitting to elucidate the positions and areas of individual diffraction peaks. The ratio of the total peak area to the areas under peaks characteristic of ‘A’ and ‘B’ type starches, respectively, were used to calculate the relative proportions of these polymorphs in smooth pea starch. These proportions were found to be 56±3% ‘A’ polymorph to 44±3% ‘B’ polymorph. A ‘C’ type pattern was constructed by using Lotus 1-2-3 to combine diffraction patterns from the crystalline portions of ‘A’ and ‘B’ type starches in the proportions given above. Polymorph patterns were obtained by manipulation of the diffraction patterns from the crystalline portions of starches using Lotus 1-2-3. An ‘A’ type pattern was obtained by subtraction of a ‘B’ type pattern from that of a ‘C’ type. Similarly, a ‘B’ type pattern was obtained by subtraction of an ‘A’ type pattern from that of a ‘C’ type.     

A characteristic pharmacological action of ‘Yang-invigorating’ Chinese tonifying herbs: Enhancement of myocardial ATP-generation capacity

In order to investigate the pharmacological basis of ‘Yang-invigorating’ action, the effect of oral treatment with the methanolic extract of ‘Yang-invigorating’ herbs on ATP-generation capacity was examined, using heart homogenates prepared from herb-pretreated mice. Tonifying (i.e., health-promoting) herbs of other functional categories were also included for comparison. The results indicated that ‘Yang-invigorating’ Chinese tonifying herbs could invariably enhance myocardial ATP-generation capacity, with the extent of stimulation varying among the herbs. In contrast, ‘Yin-nourishing’ herbs either did not stimulate or even decreased myocardial ATP-generation capacity. While ‘Qi-invigorating’ herbs produced variable effects on myocardial ATP-generation capacity, most of the ‘blood-enriching’ herbs did not cause any significant changes. The results obtained from studies using myocardial mitochondrial fractions isolated from herb-pretreated mice suggest that ‘Yang-invigorating’ herbs might speed up ATP generation by increasing mitochondrial electron transport. The ensemble of results has provided evidence for the first time to support the pharmacological basis of ‘Yang invigoration’ in Chinese medicine.     

Trunk muscle onset and cessation in golfers with and without low back pain

The knowledge of the onset and cessation timing of the paraspinal muscles that surround the lumbar spine is an important area of research for the understanding of low back pain. This study examined the timing of the erector spinae and external oblique muscle activity in a group of golfers with and without low back pain. The study compared the results of surface electromyography measurements for two groups of golfers. Twelve male golfers who had reported a mild or greater level of pain in the lower back that was experienced while playing golf were examined. A further fifteen male golfers who had reported no history of lower back pain in the previous 12 months were recruited as controls. The results showed that the low-back-pain golfers switched on their erector spinae muscle significantly in advance of the start of the backswing. This finding was not evident in the group who did not have low back pain symptoms. Low-back-pain golfers, therefore, may use the erector spinae muscle as a primary spinal stabiliser instead of the stronger deeper muscles such as transversus abdominis and multifidus. These results may have important implications for conditioning programmes for golfers with low back pain.     

Altered trunk muscle coordination during rapid trunk flexion in people in remission of recurrent low back pain

People with a history of low back pain (LBP) are at high risk to encounter additional LBP episodes. During LBP remission, altered trunk muscle control has been suggested to negatively impact spinal health. As sudden LBP onset is commonly reported during trunk flexion, the aim of the current study is to investigate whether dynamic trunk muscle recruitment is altered in LBP remission. Eleven people in remission of recurrent LBP and 14 pain free controls performed cued trunk flexion during a loaded and unloaded condition. Electromyographic activity was recorded from paraspinal (lumbar and thoracic erector spinae, latissimus dorsi, deep and superficial multifidus) and abdominal muscles (obliquus internus, externus and rectus abdominis) with surface and fine-wire electrodes. LBP participants exhibited higher levels of co-contraction of flexor/extensor muscles, lower agonistic abdominal and higher antagonistic paraspinal muscle activity than controls, both when data were analyzed in grouped and individual muscle behavior. A sub-analysis in people with unilateral LBP (n = 6) pointed to opposing changes in deep and superficial multifidus in relation to the pain side. These results suggest that dynamic trunk muscle control is modified during LBP remission, and might possibly increase spinal load and result in earlier muscle fatigue due to intensified muscle usage. These negative consequences for spinal health could possibly contribute to recurrence of LBP.     

EMG scanning in the diagnosis of chronic pain

A surface EMG diagnostic protocol was developed to assess the neuromuscular/postural contributions to pain states. The EMG activity of the right and left aspects of 11 muscle groups were monitored while the patient was in the sitting and standing positions. The diagnostic protocol was evaluated by comparing the patterns of EMG activity in four diagnostic groups: headache only, neck/shoulder/upper back pain only, low back pain only, and mixed pain states. The results suggest that (1) bilateral levels of EMG activity in the frontalis and masseter groups are of primary importance for the headache patients, (2) the discrepancy between the right and left EMG activity in the lumbar and cervical paraspinal muscle groups are of primary importance for low back pain patients, (3) position (sit/stand) may provide important diagnostic information, and (4) the data appear to support the notion of a postural disturbance as a contributing factor in low back pain.     

The effect of surgical floor mats in prolonged standing: an EMG study of the lumbar paraspinal and anterior tibialis muscles

The purpose of this study was to determine whether or not surgical floor mats affect low back and leg muscle activity during prolonged standing. The EMG activity was measured continuously using surface electrodes on the paraspinal muscles of the low back and on the anterior tibialis muscles; the subjects were normal and stood on two different surfaces. Six male subjects were each instructed to stand for two hours on a specially designed surgical floor mat and then, on a separate day, to stand for two hours on a linoleum-covered concrete surface. Six other subjects carried out the same procedure, but stood on the linoleum first. There was no difference in EMG activity obtained from the anterior tibialis muscles and paraspinal muscles of the low back when the subjects stood on the surgical mat, as compared with the linoleum-covered concrete.     

Biosynthesis of Hb in individual fetal liver bursts : γ-Chain production peaks earlier than β-chain in the erythropoietic pathway

The relative synthesis of α-, β-, ^Gγ- and ^Aγ-globin chains has been evaluated in single fetal liver bursts, which were grown in methylcellulose cultures, individually labelled with [³H]leucine and then analysed via iso-electric focusing. Well-hemoglobinized bursts demonstrate a homogeneous globin synthetic pattern, characterized by prevalent HbF (+some HbA) synthesis: thus, they apparently originate from a homogeneously programmed population of erythroid burst-forming unit (BFU-E). On day 8–9 of culture, the synthetic pattern in ‘mature’ (i.e., well-hemoglobinized) bursts has been compared with that in simultaneously-grown, ‘immature’ (i.e., poorly-hemoglobinized) colonies. These patterns have been further compared with that in ‘matured’ bursts (identified in situ as immature on day 8–9 and labelled 2–4 days later when matured). The ‘immature’ colonies showed very low levels of relative β-globin synthesis, while the ‘mature’ ones demonstrated a more elevated production of β-chain. Significantly, the ‘matured’ bursts showed a globin chain synthetic pattern similar to that of previously labelled ‘matured’ colonies. It is postulated therefore that in fetal liver (and also in adult marrow) the synthesis of γ-chain is linked to an early differentiation stage of erythroblasts, while β-globin synthesis is largely activated at a more advanced maturation stage.     

Jogging gait kinetics following fatiguing lumbar paraspinal exercise

A relationship exists between lumbar paraspinal muscle fatigue and quadriceps muscle activation. The objective of this study was to determine whether hip and knee joint moments during jogging changed following paraspinal fatiguing exercise. Fifty total subjects (25 with self-reported history of low back pain) performed fatiguing, isometric lumbar extension exercise until a shift in EMG median frequency corresponding to a mild level of muscle fatigue was observed. We compared 3-dimensional external joint moments of the hip and knee during jogging before and after lumbar paraspinal fatigue using a 10-camera motion analysis system. Reduced external knee flexion, knee adduction, knee internal rotation and hip external rotation moments and increased external knee extension moments resulted from repetitive lumbar paraspinal fatiguing exercise. Persons with a self-reported history of LBP had larger knee flexion moments than controls during jogging. Neuromuscular changes in the lower extremity occur while resisting knee and hip joint moments following isolated lumbar paraspinal exercise. Persons with a history of LBP seem to rely more heavily on quadriceps activity while jogging.     

Isolation of autofluorescent ‘aged’ human fibroblasts by flow sorting : Morphology, enzyme activity and proliferative capacity

In cultures of human fibroblasts the percentage of bright autofluorescent (AF) cells increases with increasing passage number. These autofluorescent cells were isolated using a FACS II cell sorter and compared with sorted non-fluorescent (NF) cells. The AF cells showed an increase in population doubling time (2.3-fold), cell protein (1.9-fold), and in specific activities of the lysosomal enzymes: β-hexosaminidase (4.2-fold), β-galactosidase (3.8-fold) and acid phosphatase (2.5-fold). The specific activities of two non-lysosomal enzymes glucose-6-phosphate dehydrogenase and lactate dehydrogenase had increased only slightly (1.1-fold) respectively (1.5-fold).The autofluorescence in the AF cells was restricted to small round organelles. The distribution and size of these autofluorescence granules were similar to the acid phosphatase-containing granules in the cytochemically stained cells. Electronmicroscopical examination showed that these AF cells contained a large amount of small electron-dense granules containing amorphosmophilic material. These granules which were positive for the acid phosphatase reaction, were classified as secondary lysosomes. The low percentage of the sorted AF cells which incorporate [³H]thymidine during a 24 h test period (19%) as compared with the labelling percentage of sorted NF cells (73%) from the same culture, indicate that the autofluorescent cells in a ‘young’ culture have a very limited remaining proliferative capacity. The results imply, that by flow sorting it is possible to isolate ‘aged’ cells with characteristics of ‘phase III’ cells out of non-aged fibroblast cultures.     

Sequence dependent hydration of DNA

The transitions between the different helical conformations of DNA depend on the base sequence and the ambient conditions such as humidity and counter-ion concentration. In this study energy minimization techniques have been used to locate water molecule sites around nucleotides especially those which form hydrogen bonds between two or more nucleotide atoms and thus form solvent mediated bridges. We have studied several sequences and find that those which are known not to exist in the low hydration ‘A’ form have very similar number of bridging sites in both ‘A’ and ‘B’ conformations. Those sequences which are found in the ‘A’ conformation have considerably more bridging sites in this low hydration form than in the ‘B’ conformation. Sequence related solvent effects for a given conformation have also been analysed.     

Double infection experiments with echinostomatids (Trematoda) in Lymnaea stagnalis by implantation of rediae and exposure to miracidia

1972. Double infection experiments with echinostomatids (Trematoda) in Lymnaea stagnalis by implantation of rediae and exposure to miracidia. International Journal for Parasitology, 2: 409–423. Echinostomatid species parasitizing Lymnaea stagnalis as first intermediate hosts in a South German Lake have been found present in natural double infections, but at frequencies lower than expected. Simultaneous double infection and superinfection experiments in Lymnaea stagnalis with Isthmiophora melis, Echinoparyphium aconiatum and Echinostoma revolutum were performed by redial implantation and by exposure to miracidia. All three combinations possible of these echinostomatids proved to be unstable, one species being eliminated by another ‘stronger’ one after an invariable suppression order. The degree of vigour of Isthmiophora melis in this suppression order is greater if mother rediae (macropharyngeate) are present, i.e. after miracidial invasion instead of daughter redial implantation. Snails parasitized by rediae of a ‘weak’ type could be superinfected by implantation of rediae of a ‘strong’ type, but not if the first (‘weak’) infection had reached the stage of shedding cercariae. Superinfection by implantation of Echinoparyphium aconiatum rediae (‘strong’ type) was not successful when the first infection consisted of sporocysts of plagiorchiids, or of Apatemon sp. (Strigeidae) that had reached the stage of shedding cercariae.     

The regulatory function of the myosin light chains

In molluscan muscles, calcium regulation is mediated by ‘regulatory’ light chains associated with the myosin heads. This type of ‘regulatory’ light chain appears to be present in all myosins, regardless of whether the myosin contains light chain linked calcium regulation. Although they appear to be ‘structurally’ related, differences in their calcium binding abilities imply that these regulatory light chains may play quite distinct functions in their respective myosins.     

'P' solubilization potential of plant growth promoting Pseudomonas mutants at low temperature

Pseudomonas fluorescens strain GRS₁, PRS₉ and their cold tolerant mutants were examined for their tricalcium phosphate (TCP) solubilizing activity in NBRIP (broth) media at 10°C and 25°C. Invariably, all the cold tolerant mutants of GRS₁ and PRS₉ were found more efficient than their respective wild type counterparts for ‘P’ solubilization activity at 10°C as compared to 25°C. ‘P’ solubilization potential of CRM was found maximum among all the strains followed by CRPF₆ and CRPF₄. To the best of out knowledge, this is the first report regarding low temperature ‘P’ solubilization activity.     

Trunk muscle responses to suddenly applied loads: Do individuals who develop discomfort during prolonged standing respond differently?

Individuals with low back pain or injury (LBP/LBI) have been shown to display altered muscle responses to trunk perturbations; however it is unclear whether these observations are a cause or a result of the LBP/LBI. In this study, a 6.78 kg load was suddenly applied to the hands to perturb the trunk prior to and following a 2-h standing period, during which trunk and hip electromyography (EMG) and centre of pressure (CoP) at the feet were recorded. Seven of the 13 participants developed substantial low back discomfort (LBDiscomfort) during the standing period. These individuals, both pre- and post-standing, showed a greater average number of responsive extensor muscles (3.8 compared to 3.1 in those who did not develop discomfort) and a greater occurrence of extensor muscle response (95–100% of trials) as compared to those who did not develop LBDiscomfort (73–86% of trials). Also, after discomfort developed, these individuals displayed an increased response in their abdominal muscles. This overall increase in trunk musculature activity could either be detrimental by potentially increasing spinal loading leading to LBDiscomfort, or beneficial in that this increased musculature responsiveness may reduce one’s likelihood of developing a future LBI through a pathway of increased spine stability. In either case, these responses indicate motor control characteristics that can distinguish the likelihood of an individual developing LBDiscomfort during common tasks such as prolonged standing.     

Intracellular signalling proteins as ‘smart’ agents in parallel distributed processes

In eucaryotic organisms, responses to external signals are mediated by a repertoire of intracellular signalling pathways that ultimately bring about the activation/inactivation of protein kinases and/or protein phosphatases. Until relatively recently, little thought had been given to the intracellular distribution of the components of these signalling pathways. However, experimental evidence from a diverse range of organisms indicates that rather than being freely distributed, many of the protein components of signalling cascades show a significant degree of spatial organisation. Here, we briefly review the roles of ‘anchor’, ‘scaffold’ and ‘adaptor’ proteins in the organisation and functioning of intracellular signalling pathways. We then consider some of the parallel distributed processing capacities of these adaptive systems. We focus on signalling proteins-both as individual ‘devices’ (agents) and as ‘networks’ (ecologies) of parallel processes. Signalling proteins are described as ‘smart thermodynamic machines’ which satisfy ‘gluing’ (functorial) roles in the information economy of the cell. This combines two information-processing views of signalling proteins. Individually, they show ‘cognitive’ capacities and collectively they integrate (cohere) cellular processes. We exploit these views by drawing comparisons between signalling proteins and verbs. This text/dialogical metaphor also helps refine our view of signalling proteins as context-sensitive information processing agents.     

Paraspinal muscle vibration alters dynamic motion of the trunk

Loss in dynamic stability of the low back has been identified as a potential factor in the etiology of low back injuries. A number of factors are important in the ability of a person to maintain an upright trunk posture including the preparatory stiffness of the trunk and the magnitude and timing of the neuromotor response. A neuromotor response requires appropriate sensing of joint motion. In this research, the role of this sensory ability in dynamic performance of the trunk was examined using a simple pendulum model of the trunk with neuromotor feedback. An increased sensory threshold was found to lead to increased torso flexion and increased delay in neuromotor response. This was confirmed experimentally using paraspinal muscle vibration which is known to alter proprioception of the muscle spindle organs. Before, during and after exposure to bilateral, paraspinal muscle vibration for 20 minutes, the dynamic response of subjects to an unexpected torso flexion load was examined. Subjects were found to have a 19.5% slower time to peak muscle activity and a 16.1% greater torso flexion during exposure to paraspinal muscle vibration. Torso flexion remained significantly increased after vibration exposure relative to before exposure. These results suggest that the neuromotor response plays an important role in trunk dynamics. Loss in sensitivity of the sensory system can have a detrimental effect on trunk dynamics, increasing delays in neuromotor response and increasing the motion of the trunk in response to an unexpected load.     

Application of skinned single muscle fibres to determine myofilament function in ageing and disease

The chemically skinned fibre is a suitable preparation to determine whether alterations in myofilament function contribute to muscle dysfunction during ageing and disorders such as chronic obstructive pulmonary disease (COPD). In this preparation the sarcolemma is chemically permeabilized and the myofilament lattice kept intact, functioning under controlled near-physiological conditions. As force generating capacity is an important determinant of muscle function and is related to fibre crosssectional area (FCSA), we compared several methods employed by researchers to determine FCSA. Specific tension, force divided by FCSA, has a co-efficient of variation of 27%, 37%, or 30% when the FCSA was measured from the width and depth assuming an elliptical circumference, the width assuming a circular circumference, and the width while the fibre was suspended in the air, respectively. The last method showed the closest relation with the FCSA in histological sections. The velocity of maximal unloaded shortening (V(0)) varied with fibre type, with fibres expressing the Beta/slow (type I) myosin heavy chain (MyHC) isoform being the slowest and fibres expressing the IIb MyHC isoform the fastest. While muscle weakness experienced after surgery could not be explained by changes in specific tension or FCSA of individual fibres, the preparation revealed significant changes in myofilament function during ageing and COPD.     

Transitions between cooperative and non-cooperative responding in the 'Pigeon's Dilemma'

Two pigeons played Iterated Prisoner's Dilemma (IPD) against a simulated opponent pre-programmed to play ‘Tit-For-Tat’ (TFT) and ‘Random’ (RND) strategies. Each pigeon received differential amounts of access to food following choices of either ‘cooperate’ or ‘defect’ on a trial. After 1000 trials against TFT and 500 trials against RND, results indicated that choice allocation was optimal when the birds played against RND but was sub-optimal when the birds played against TFT. In order to determine why the pigeons responded suboptimally against TFT, a trial-by-trial analysis of the data was conducted. The analysis revealed that once a pigeon had received the ‘Sucker's’ payoff (S), it was more likely to defect and receive the ‘Temptation’ payoff (T) than to cooperate and receive the ‘Reward’ (R) payoff. Local reinforcement contingencies appear to determine suboptimal responding against TFT in the iterated Pigeon's Dilemma.